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TCA沉淀法定量DNA

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摘要:Mix5mlofthesamplewith100mlSalmonspermDNA(50mgin20mMEDTA)。Add5mLicecold10%Trichloroaceticacid(TCA)。Washthefilter6timeswith5mLice-cold10%TCA。ThesecondfiltermeasuresradioactivityincorporatedintoDNAfragmentsgreaterthan20nucleotidesinlength。...

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1. Dilute radioactive sample to a 100 ml volume

2. Spot 5 ml of the sample onto the center of a 2.4 cm Whatman GF/C glass-fiber disc.

3. Mix 5 ml of the sample with 100 ml Salmon sperm DNA (50 mg in 20 mM EDTA).

4. Add 5 mL ice cold 10% Trichloroacetic acid (TCA). Mix well and incubate on ice for 15 min.

5. Filter the solution through a separate GF/C glass-fiber disc.

6. Wash the filter 6 times with 5 mL ice-cold 10% TCA.

7. Wash filter once with 5 mL 95% Ethanol.

8. Dry both filter under a heat lamp.

9. When dry, count each in a scintillation counter.

10. The first filter measures total radioactivity. The second filter measures radioactivity incorporated into DNA fragments greater than 20 nucleotides in length.

相关产品链接:

试剂库 >>核酸检测 >> 核酸定量检测

试剂库 >>PCR/RT-PCR/qPCR >>定量PCR试剂

仪器库 >> 分子生物实验仪器 >>紫外设备

仪器库 >> 影像系统 >>凝胶成像系统

仪器库 >> 测读系统>>分光光度计

作者: 2008-2-3
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